Genome-Wide ENU Mutagenesis in Combination with High Density SNP Analysis and Exome Sequencing Provides Rapid Identification of Novel Mouse Models of Developmental Disease
Date
2013-03-01
Authors
Caruana, Georgina
Farlie, Peter G
Hart, Adam H
Bagheri-Fam, Stefan
Wallace, Megan J
Dobbie, Michael S
Gordon, Christopher T
Miller, Kerry A
Whittle, Belinda
Abud, Helen E
Journal Title
Journal ISSN
Volume Title
Publisher
Public Library of Science
Abstract
BACKGROUND Mice harbouring gene mutations that cause phenotypic abnormalities during organogenesis are invaluable tools for linking gene function to normal development and human disorders. To generate mouse models harbouring novel alleles that are involved in organogenesis we conducted a phenotype-driven, genome-wide mutagenesis screen in mice using the mutagen N-ethyl-N-nitrosourea (ENU). METHODOLOGY/PRINCIPAL FINDINGS ENU was injected into male C57BL/6 mice and the mutations transmitted through the germ-line. ENU-induced mutations were bred to homozygosity and G3 embryos screened at embryonic day (E) 13.5 and E18.5 for abnormalities in limb and craniofacial structures, skin, blood, vasculature, lungs, gut, kidneys, ureters and gonads. From 52 pedigrees screened 15 were detected with anomalies in one or more of the structures/organs screened. Using single nucleotide polymorphism (SNP)-based linkage analysis in conjunction with candidate gene or next-generation sequencing (NGS) we identified novel recessive alleles for Fras1, Ift140 and Lig1. CONCLUSIONS/SIGNIFICANCE In this study we have generated mouse models in which the anomalies closely mimic those seen in human disorders. The association between novel mutant alleles and phenotypes will lead to a better understanding of gene function in normal development and establish how their dysfunction causes human anomalies and disease.
Description
Keywords
alleles, animals, congenital abnormalities, dna ligases, ethylnitrosourea, exome, extracellular matrix proteins, female, genome-wide association study, genotype, germ-line mutation, high-throughput nucleotide sequencing, homozygote, left-right determination factors, male, mice, mice, inbred c57bl, mutagenesis, mutagens, phenotype, disease models, animal, polymorphism, single nucleotide
Citation
Collections
Source
PLoS ONE
Type
Journal article
Book Title
Entity type
Access Statement
License Rights
Restricted until
Downloads
File
Description
Published Version